快速細菌藥物敏感性試驗（AST）是及時治療感染和預防細菌抗藥性的最有效策略之一。然而，抗生素的濫用使得抗藥性細菌日漸增多，成為人類健康的一大威脅。為了突破此困境，透過本實驗室之核心技術-布朗運動為基礎，開發一光擴散儀觀測修飾微珠和細菌之間的相互作用。根據斯托克斯-愛因斯坦方程式(Stoke-Einstein equation)，擴散率與微珠直徑成反比，若擴散率下降表示細菌數目增加並與微珠結合，基於微珠擴散率的與時間的趨勢變化，進一步得出細菌藥敏試驗的結果。所有實驗能在四小時內完成藥敏試驗(包含檢體前處理)。目前我們正著手發展性能更為強大的光擴散儀，在本研究中，我們將設計、製作以及分析一多孔微晶片之特徵以用於搭載在該擴散儀上。在設計上，此多孔微晶片具有多檢測腔室，有著更高的檢測數量 (300 個腔室)，以利我們進行大規模的藥敏試驗，並透過模擬軟體觀察結構擴散情形。填充方式上僅僅靠一支特殊設計的針筒來填充試片，此方法不僅不依靠外部驅動力，並且操作簡單，醫療人員不用經過特殊訓練即可上手。抗生素封存上我們進行了光譜以及生物培養測試，確信了凍乾將會是以後製造微晶片時，藥物封存方式的首選。此多孔微晶片在經驗證效能後，未來我們期許能具有大量製造並應用之潛力。除此之外，藉由相同技術，我們同時也開發用於快速免疫分析之光擴散儀，分子抗原的接枝使用三明治法進行兩次專一性的辨認，並以腫瘤壞死因子α作為首先的生物標靶進行實測。由實驗得出之結果，此光擴散儀已經能達到從量測到分析(<10 min)、輕便可攜式(< 6 kg)、低樣本量(~20 μL)以及檢測最低濃度(< 6 ng/mL)等優點，我們預期在未來此免疫分析儀將能成為定點照護(Point-of-care testing. POCT)的一項強力工具。

The rapid antimicrobial susceptibility testing (AST) is the most effective strategies in the timely treatment of infections and preventing bacterial drug resistance. However, the abuse of antibiotics leads to increase the number of resistant bacteria which has be-come a serious threat to human health. To deal with this ordeal, we developed an optical diffusometry which applied our core-technology, Brownian motion, in our lab to inves-tigate the interactions between antibiotic-treated bacteria and functionalized microbeads. According to the Stokes- Ein-stein equation, the diffusivity is inversely proportional to the microbead diameter. The diffusivity decreased when the microbead diameter in-creased. When the number of bacteria increased, the microbead would conjugating with bacteria, hence resulting in a time-dependent diffusivity change. Based on the dependent trend between time and microbead diffusivity, we could obtain the result about antimi-crobial susceptibility within 4 h including pre-preparation of sample. Now, we are con-tributing to develop a more advanced optical diffusometry. In this thesis, we would de-sign, fabricate, and characterize of multi-well microchip for new optical diffusometry. It is more convenience to do large scale test on the new microchip because it has more de-tection chamber in structure (300 wells). Load up the microchip only depended by a special syringe without external driving force. This way is simple and without training to operate for medical personnel. In the antibiotic storage of microchip, we ensure that lyophilization is the best way to maintain the effect of drug after conducting the test of spectrum and biological activity. Through verification the performance of this micro-chip, we believe that it has great potential on mass production for medical application. Furthermore, we simultaneously developed a rapid and portable optical diffusometry for immunoassay. We used the sandwich Enzyme-linked immunosorbent assay to perform twice specific identifications in conjugate antigen. The Tumor Necrosis Factor alpha which is related a lot of dis-ease as a biomarker would be first detected in this thesis. Observing the result from experiment, our optical diffusometry can achieve rapid anal-ysis(< 10 min), light and portable (< 6 kg), low sample volume (~20 mL) and quantita-tive analysis at a low concentration (<6 ng/mL). We proved that the benchtop platform can be a powerful tool in the point-of-care test of immunoassay in the near future.

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